Fig. 2

Knocking out STC1 in CAFs attenuated their ability to promote HCC stemness. A The selection of shRNA sequences and the regulation of STC1 in CAFs. B A sphere formation assay was used to analyze the self-renewal ability of HCC cells. MHCC-97H and SNU-398 cells were treated with DMEM, CAF-shcontrol-CM, CAF-shSTC1-1-CM, or CAF-shSTC1-2-CM. Scale bar, 100 μm. C HCC cells were treated with different conditioned medium of CAFs for 48 h, and their viability was assessed after 24 h of sorafenib treatment. D A colony formation assay was performed in the indicated cells. E Representative images of transwell migration and invasion assays of different groups of MHCC-97H cells. Scale bar, 100 μm. F Western blot analysis of the expression of cancer stemness markers in the indicated HCC cells. G HCC cells with or without the indicated CAFs were transplanted into the livers of nude mice. Representative bioluminescence images of each group are shown. For the statistical analysis, ns, no significance, *P < 0.05, **P < 0.01, and ***P < 0.001, t test